Using patterned supported lipid membranes to investigate the role of receptor organization in intercellular signaling

Nair, Pradeep M.; Salaita, Khalid; Petit, Rebecca S.; Groves, Jay T.

doi:10.1038/nprot.2011.302

Cited by 91 publications

(90 citation statements)

References 45 publications

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“…27 The lift-off process resist patterns generated with electron-beam lithography typically produce Cr barriers a few nanometers high and a hundred nanometers wide. These barriers are used to engineer the spatial organization of cell membrane receptors for interfacing with living cells.…”

Section: Formation Of Lipid-nanostructure Hybrids and Their Structurementioning

confidence: 99%

Lipid-nanostructure hybrids and their applications in nanobiotechnology

Lee

Nam

2013

NPG Asia Mater

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Cell membranes contain a variety of lipids and functional proteins that offer active platforms that organize the membrane components into functional assemblies and perform biologically important reactions. The dynamic and complex nature of the membranes makes them attractive materials, but at the same time, researchers report substantial experimental uncertainty in controlling the membrane reactions and extracting valuable information. The fascinating new structures and properties of nanomaterials could be utilized in addressing aforementioned issues, and the design and synthesis of lipid-nanostructure hybrids could be beneficial to the research areas in lipid-membrane biotechnology and nanobiotechnology. These hybrid structures possess dimensions that are comparable to that of biological molecules and structures and physicochemical properties that arise from both lipids and nanomaterials. Therefore, lipid-nanostructure hybrids offer additional options for control of the synthesized structures, provide new insight in understanding nanostructures and biological systems and allow the mimicking of functional subcellular membrane components and monitoring of the membrane-associated reactions in a highly sensitive and controllable manner. In this review, we present recent advances in the synthesis of various lipid-nanostructure hybrids and the application of these structures in biotechnology and nanotechnology. We further describe the scientific and practical applications of lipid-nanostructure hybrids for detecting membrane-targeting molecules, interfacing nanostructures with live cells and creating membrane-mimicking platforms to investigate various intercellular processes.

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Section: Formation Of Lipid-nanostructure Hybrids and Their Structurementioning

confidence: 99%

Lipid-nanostructure hybrids and their applications in nanobiotechnology

Lee

Nam

2013

NPG Asia Mater

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“…Importantly, for the study of membrane-associated biological processes, model membranes enjoy the advantage of self-assembly formation [31,34], which permits facile adjustment of system parameters. As such, bottom-up design has enabled the development of model membrane systems for probing a wide range of biological processes such as cell adhesion, membrane fusion, and cellular signaling, and there are many excellent reviews already in the field [35][36][37][38][39]. However, there has been limited demonstration thus far of how knowledge gained from these approaches can be translated into clinically impactful biomedical applications.…”

Section: Introductionmentioning

confidence: 99%

Model Membrane Platforms for Biomedicine: Case Study on Antiviral Drug Development

Jackman

Cho

2012

Biointerphases

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As one of the most important interfaces in cellular systems, biological membranes have essential functions in many activities such as cellular protection and signaling. Beyond their direct functions, they also serve as scaffolds to support the association of proteins involved in structural support, adhesion, and transport. Unfortunately, biological processes sometimes malfunction and require therapeutic intervention. For those processes which occur within or upon membranes, it is oftentimes difficult to study the mechanism in a biologically relevant, membranous environment. Therefore, the identification of direct therapeutic targets is challenging. In order to overcome this barrier, engineering strategies offer a new approach to interrogate biological activities at membrane interfaces by analyzing them through the principles of the interfacial sciences. Since membranes are complex biological interfaces, the development of simplified model systems which mimic important properties of membranes can enable fundamental characterization of interaction parameters for such processes. We have selected the hepatitis C virus (HCV) as a model viral pathogen to demonstrate how model membrane platforms can aid antiviral drug discovery and development. Responsible for generating the genomic diversity that makes treating HCV infection so difficult, viral replication represents an ideal step in the virus life cycle for therapeutic intervention. To target HCV genome replication, the interaction of viral proteins with model membrane platforms has served as a useful strategy for target identification and characterization. In this review article, we demonstrate how engineering approaches have led to the discovery of a new functional activity encoded within the HCV nonstructural 5A protein. Specifically, its N-terminal amphipathic, α-helix (AH) can rupture lipid vesicles in a size-dependent manner. While this activity has a number of exciting biotechnology and biomedical applications, arguably the most promising one is in antiviral medicine. Based on the similarities between lipid vesicles and the lipid envelopes of virus particles, experimental findings from model membrane platforms led to the prediction that a range of medically important viruses might be susceptible to rupturing treatment with synthetic AH peptide. This hypothesis was tested and validated by molecular virology studies. Broad-spectrum antiviral activity of the AH peptide has been identified against HCV, HIV, herpes simplex virus, and dengue virus, and many more deadly pathogens. As a result, the AH peptide is the first in class of broad-spectrum, lipid envelope-rupturing antiviral agents, and has entered the drug pipeline. In summary, engineering strategies break down complex biological systems into simplified biomimetic models that recapitulate the most important parameters. This approach is particularly advantageous for membrane-associated biological processes because model membrane platforms provide more direct characterization of target interactions than is possible with other methods. Consequently, model membrane platforms hold great promise for solving important biomedical problems and speeding up the translation of biological knowledge into clinical applications.

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“…However, while these methods are powerful and applicable to biomolecular patterning, their resolution is essentially limited to that of conventional DPN, which is significantly larger than most soluble protein molecules that are generally smaller than 10 nm in diameter. There are also some examples of using traditional methods such as electron beam lithography to prepare small collections of particles that can support individual protein attachment; the ability to control the placement of biomolecules with this degree of resolution and precision over large areas remains a challenge for current nanolithographic processes (16)(17)(18). Indeed, the conventional methods are expensive, inherently low throughput, and difficult to implement on the sub-10-nm scale.…”

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confidence: 99%

Single-molecule protein arrays enabled by scanning probe block copolymer lithography

Chai¹,

Wong²,

Giam

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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The ability to control the placement of individual protein molecules on surfaces could enable advances in a wide range of areas, from the development of nanoscale biomolecular devices to fundamental studies in cell biology. Such control, however, remains a challenge in nanobiotechnology due to the limitations of current lithographic techniques. Herein we report an approach that combines scanning probe block copolymer lithography with site-selective immobilization strategies to create arrays of proteins down to the single-molecule level with arbitrary pattern control. Scanning probe block copolymer lithography was used to synthesize individual sub-10-nm single crystal gold nanoparticles that can act as scaffolds for the adsorption of functionalized alkylthiol monolayers, which facilitate the immobilization of specific proteins. The number of protein molecules that adsorb onto the nanoparticles is dependent upon particle size; when the particle size approaches the dimensions of a protein molecule, each particle can support a single protein. This was demonstrated with both gold nanoparticle and quantum dot labeling coupled with transmission electron microscopy imaging experiments. The immobilized proteins remain bioactive, as evidenced by enzymatic assays and antigen-antibody binding experiments. Importantly, this approach to generate single-biomolecule arrays is, in principle, applicable to many parallelized cantilever and cantilever-free scanning probe molecular printing methods.dip-pen nanolithography | scanning probe lithography | single molecule array | gold nanoparticles | protein nanoarray P rotein immobilization on solid substrates with nanoscale control has been utilized in a variety of applications, including chip-based bioassays (1), proteomics (2, 3), drug discovery (3), and cellular biology studies (4). In cellular biology research, the ability to fabricate protein nanostructures on surfaces has enabled the study of many basic cellular functions including growth, signaling, and differentiation (4-7). For combinatorial molecular biology, the miniaturization of protein nanoarrays allows for smaller and higher density chips and the need for smaller sample volumes; in certain cases, this can translate into diagnostic systems with higher sensitivity and the ability to track disease and biological processes more efficiently (2, 3). The ability to site-specifically isolate single biomolecules can also facilitate molecular level studies of such structures (8, 9). Therefore, being able to nanofabricate biomolecular features at a resolution of 10 nm or less is of significant interest because this length scale approaches the dimensions of single protein molecules and offers an opportunity to address many previously unexplored biological phenomena.The use of dip-pen nanolithography (DPN) (4, 10) for the generation of arrays of biomolecules either by a direct deposition of proteins (11-13) or indirect methods (14, 15), through DPN writing of patterns followed by capture of proteins onto the patterns, has been widel...

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Using patterned supported lipid membranes to investigate the role of receptor organization in intercellular signaling

Cited by 91 publications

References 45 publications

Lipid-nanostructure hybrids and their applications in nanobiotechnology

Lipid-nanostructure hybrids and their applications in nanobiotechnology

Model Membrane Platforms for Biomedicine: Case Study on Antiviral Drug Development

Single-molecule protein arrays enabled by scanning probe block copolymer lithography

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